Journal: Nature Communications

Article Title: AAV-delivered muscone-induced transgene system for treating chronic diseases in mice via inhalation

doi: 10.1038/s41467-024-45383-z

Figure Lengend Snippet: a Schematic representing the genetic configuration of the AAV vector used for AAV MUSE iteration. b Schematic representation of the experimental procedure for AAV MUSE -mediated transgene expression in mouse lungs. c AAV MUSE -mediated luciferase expression for 20 weeks in mouse lungs. Eight-week-old female BALB/c mice were transduced with an AAV MUSE iteration comprising AAV2/lung-pWX126 (ITR-P SV40 -MOR215-1-pA-ITR, 5 × 10 11 vg) and AAV2/6-pWX342 (ITR-P SV40 -RTP1S::pA-luciferase-P CRE -ITR, 5 × 10 11 vg) via tail vein. Two weeks after the AAV injection, AAV MUSE -transduced mice were exposed to nebulized muscone for 4 h once every six weeks or exposed to a vehicle (a mixture of castor oil and ddH 2 O), and bioluminescence imaging was performed using an in vivo imaging system every 6 weeks. d Quantification of luciferase expression in the lung based on the bioluminescence IVIS imaging shown in ( c ). Data in ( d ) are presented as means ± SEM ( n = 4 mice). ** P < 0.01, *** P < 0.001. P values were obtained from two-tailed unpaired t tests. Source data are provided as a file.

Article Snippet: AAV (serotype 2/6) carrying pWX342 (ITR-P SV40 -RTP1S::pA-luciferase-P CRE -ITR) or pWX345 (ITR-P SV40 -RTP1S::pA-ΔmIL-4-P CRE -ITR) and AAV (serotype 2/lung) carrying pWX126, pWX127, or pWX158 were produced by Shanghai OBiO Technology.

Techniques: Plasmid Preparation, Expressing, Luciferase, Transduction, Injection, Imaging, In Vivo Imaging, Two Tailed Test

Journal: Sensors (Basel, Switzerland)

Article Title: Human Olfactory Receptor Sensor for Odor Reconstitution

doi: 10.3390/s23136164

Figure Lengend Snippet: Endpoint measurement of the binding affinity of odor molecules to OR expressed on heterologous cells. To promote OR expression on the surface of heterologous cells, a signal peptide (Rho-tag) is added to the N -terminus and the OR chaperones RTPs (RTP1, RTP1S, RTP2) and REEP1 and Gα protein chaperone Ric-8B are coexpressed. Binding of the odor molecule causes OR to activate adenylate cyclase (AC) via trimeric G proteins to produce cAMP. The amount of cAMP is assayed by endpoint measurement of chemiluminescence using GloSensor TM and is determined as the binding affinity between the odor molecules and OR.

Article Snippet: Matsunami et al. found that human embryonic kidney-derived HEK293 cells introduced with the chaperones RTP1 (receptor-transporting protein 1), RTP2, and REEP1 (receptor expression-enhancing protein 1), also called Hana3A cells, greatly improved cell surface expression of ORs with an N -terminal Rho-tag (rhodopsin-derived signal peptide) [ ] ( ).

Techniques: Binding Assay, Expressing